JPH0149156B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to novel dibenzo-p-dioxin derivatives. The dibenzo-P-dioxin derivative of the present invention is a novel compound that has not been described in any literature and is represented by the following general formula (1). [In the formula, R is the same or different and represents a hydrogen atom, a lower alkyl group, a lower alkenyl group, a phenyl lower alkyl group, or a lower alkanoyl group. ] The present inventors have discovered the seaweed Ecklonia kurome (Ecklonia kurome).
Okamura) has been intensively researched on the extract. The present inventors discovered the presence of a compound in the extract that has an inhibitory effect on plasmin inhibitors, and succeeded in extracting and isolating this compound, thereby completing the present invention. In this specification, examples of lower alkyl groups include methyl, ethyl, propyl, isopropyl, butyl, tert-butyl, pentyl, and hexyl groups. Examples of lower alkenyl groups include vinyl, allyl, 1-butenyl, 2-butenyl, 1-methyl-allyl, 2-pentenyl, and 2-hexenyl groups. Examples of phenyl lower alkyl groups include benzyl, 2-phenylethyl, 1-phenylethyl, 3-phenylpropyl, 4-phenylbutyl, 1,1-dimethyl-2-phenylethyl, 5
-phenylpentyl, 6-phenylhexyl, 2
-methyl-3-phenylpropyl group and the like. Examples of the lower alkanoyl group include formyl, acetyl, propionyl, butyryl, isobutyryl, pentanoyl, tert-butylcarbonyl, and hexanoyl groups. The compound of the present invention represented by the above general formula (1) is:
Alpha-2 plasmin inhibitor (α ₂ −
plasmininhibitor) and alpha 2-macroglobulin (α ₂ -macroglobulin). Various biological reactions such as blood coagulation and fibrinolytic phenomena (fibrinolysis) are mediated by various proteolytic enzymes, but the functions of these proteases are controlled by inhibitory protein proteins present in the body. There is. Among these inhibitor proteins, the above-mentioned plasmin inhibitor has a strong inhibitory effect on plasmin related to the fibrinolytic system, and is known to act as an inhibitor of the fibrinolytic system. Furthermore, it is known that the above-mentioned plasmin inhibitors strongly inhibit the generated plasmin in the attempt to increase fibrinolysis by activating (producing) plasmin by administering urokinase or streptokinase, which are currently used as thrombolytic agents. There is. Therefore, there is a strong desire in this field to develop a drug that can increase fibrinolysis by blocking the action of these plasmin inhibitors in the blood [Nobuo Aoki et al.: In vivo protease inhibitors; Vol. 14, No. 6, pp. 1099-1111 (1977), Tamotsu Matsuda: Hypercoagulable state; Small molecule dextran urokinase bibliography, pp. 1-15, edited and published by Otsuka Pharmaceutical Co., Ltd., January 10, 1977. Publication reference]. The compound of the present invention represented by the above general formula (1) is:
As is clear from the pharmacological test results described below, it has strong anti-plasmin inhibitor activity and is therefore useful as a prophylactic and therapeutic agent for thrombosis due to increased fibrinolysis, and also as an adjunct to conventional thrombosis treatment agents. is also useful. The compound of the present invention is produced, for example, according to the method shown below. Among the compounds of the present invention, the compound represented by the following formula 1a is extracted and isolated from, for example, the seaweed Kurome in the following manner. That is, first, the seaweed Kurome is extracted using a common polar solvent such as methanol, ethanol, isopropanol, their hydrous alcohols, and ethyl acetate, and this extract is concentrated under reduced pressure to obtain a primary extract. From the primary extract, the general formula
The method for collecting compound 1a is not particularly limited, and any of various known methods utilizing physical and chemical properties can be employed. For example, the solubility difference with impurities, common adsorbents such as activated carbon, XAD-2,
This can be carried out by methods that utilize differences in adsorption affinity for silica gel, ion exchange resins, Cephadex, etc., differences in distribution rates between two liquid phases, etc., or by a combination of these methods. More specifically, ethyl acetate,
Extraction is performed using a solvent such as chloroform or ether, and then this extract is concentrated under reduced pressure, and then subjected to Celite column chromatography, Cephadex LH-20 column chromatography, etc., and an appropriate solvent such as ethyl ether, acetone, methanol, etc. A compound of formula 1a can be obtained by elution at . Among the compounds of the present invention, compounds other than the compound of formula 1a above are produced, for example, by the method shown below. Reaction formula 1 [In the formula, R ¹ and R ² are the same or different and represent a hydrogen atom, a lower alkyl group, a lower alkenyl group, a phenyl lower alkyl group, or a lower alkanoyl group. However, at least one of each R ¹ shall represent a hydrogen atom, and all R ² shall not represent a hydrogen atom. ] For the acylation of the compound of formula 1b, a wide range of reaction conditions for ordinary acylation reactions can be employed. A wide variety of conventionally known acylating agents can be used, including lower alkanoic acids such as acetic acid and propionic acid, lower alkanoic anhydrides such as acetic anhydride, and lower alkanoic acid halides such as acetyl chloride and propionyl bromide. be able to. The amount of such an acylating agent to be used is not particularly limited and can be appropriately selected within a wide range, but it is usually at least an equimolar amount, preferably an equimolar amount, based on the raw material to replace one hydroxyl group of the compound of formula 1b. It is preferable to use a molar to 10 times molar amount. When using an acid anhydride or an acid halide as an acylating agent, the acylation reaction is preferably carried out in the presence of a basic compound. Specifically, basic compounds include metallic sodium,
Examples include alkali metals such as metallic potassium, hydroxides, carbonates, or bicarbonates of these alkali metals, alkyllithiums, and tertiary amine compounds such as pyridine, N-methylpiperidine, and triethylamine. Furthermore, when lower alkanoic acids are used as the acylating agent, the acylation reaction is preferably carried out in the presence of a dehydrating agent. Specific examples of the dehydrating agent include mineral acids such as sulfuric acid and hydrochloric acid, and sulfonic acids such as para-toluenesulfonic acid, benzenesulfonic acid and ethanesulfonic acid. The acylation reaction is carried out either without a solvent or in a solvent. Examples of solvents include ketones such as acetone and methyl ethyl ketone, ethers such as ether, dioxane, and tetrahydrofuran, aromatic hydrocarbons such as benzene, toluene, and xylene, water, pyridine, dimethylformamide, dimethyl sulfoxide, and hexamethyl phosphoric triamide. ,1,2
-dimethoxyethane and the like. The reaction proceeds either under cooling, at room temperature, or under heating, but it is usually preferable to carry out the reaction at 0 to 150°C. In particular, when an acid anhydride or an acid halide is used as the acylating agent, the reaction is preferably carried out at 0 to 80°C, and when a lower alkanoic acid is used as the acylating agent, the reaction is preferably carried out at 50 to 120°C. is preferable. The above reaction is generally completed in about 0.5 to 24 hours. For the alkylation of the compound of formula 1b, a wide range of reaction conditions for conventional alkylation reactions can be employed. For example, the expression
For alkylation of compound 1b, a suitable alkylating agent is used instead of the acylating agent and the reaction temperature is -
The reaction conditions for the above acylation reaction can be applied as they are, except that the temperature is 80 to 150°C. As the alkylating agent, a wide variety of conventionally known alkylating agents can be used, for example, the general formula R ^{2 '} Indicates a halogen atom. ], diazoalkanes such as diazomethane, dialkyl sulfates such as dimethyl sulfate, Meerwein reagents such as trimethyl oxonium fluoroborate, and the like. Specifically, the compounds of general formula (2) include methyl chloride, methyl bromide, ethyl iodide, 1-propyl bromide, 1-pentyl iodide, 1-allyl iodide, 1-butenyl bromide, Examples include benzyl, 1-bromide-2-phenylethyl, 1-iodide-2-methyl-3-phenylpropyl, and the like. When using diazoalkanes or Meerwein's reagent as an alkylating agent, it is preferable to carry out the reaction at -80 to 50°C. Furthermore, when using diazoalkanes as an alkylating agent, tetrafluoroboron hydride, p-toluenesulfonic acid, BF ₃ O
Compounds such as (C ₂ H ₅ ) ₂ may also be present. A compound of formula 1c is thus prepared. In the above alkylation, if the compound of general formula (2) is used as an alkylating agent, an alkali metal hydride is present in the reaction system, and the reaction is carried out at -78 to -15°C, R ² becomes dibenzo - Compounds introduced at the 9-position of the p-dioxy skeleton (9-substituted product), compounds introduced at the 4-position and 9-position (4,9-disubstituted product), and compounds introduced at the 2-position, 4-position, and 9-position The introduced compound (2,4,9-trisubstituted product) can be advantageously produced. Also, among the compounds of general formula (2), the benzyl halide is 1 to 3
If about twice the molar amount is used, alkyllithium is present in the reaction system, and the reaction is carried out at about -78 to -25℃,
2,4-disubstituted products can be advantageously prepared. Formulating diazoalkanes as alkylating agents
Use about 3 to 6 times the molar amount of compound 1b,
If tetrafluoroborohydride is present in the reaction system and the reaction is carried out at about -78°C to room temperature, 9-substituted products, 4,9-disubstituted products and 2,4,9-trisubstituted products can be advantageously produced. can be manufactured. Furthermore, 9-substituted, 4,9
-disubstituted and 2,4,9-trisubstituted products can be advantageously prepared. In this case, the diazoalkanes have the formula 1b
If it is used in a large excess amount (at least 3 times the mole or more) with respect to the compound, a 2,4,9-trisubstituted product can be obtained as the main component. Furthermore, an excess amount (usually 1.5 to 10 times the molar amount) of Mairwein reagent with respect to the compound of formula 1b was used as an alkylating agent, and −78
9-substituted products, 4,9-disubstituted products, and 2,4,9-trisubstituted products can be advantageously produced by carrying out the reaction at about ℃ to room temperature. The compound of Formula 1b used as a starting material in Reaction Formula-1 above is produced, for example, by the method shown below. Reaction formula 2 [In the formula, each R ³ is the same or different and represents a hydrogen atom or a lower alkanoyl group. However, at least one of each R ³ represents a lower alkanoyl group. ] The acylation of formula 1a' can be carried out under the same reaction conditions as the acylation of formula 1b above. Reaction formula 2 [In the formula, each R ⁴ is the same or different and represents a hydrogen atom, a lower alkyl group, a lower alkenyl group, or a phenyl lower alkyl group. However, at least one of each R ⁴ represents a lower alkyl group, a lower alkenyl group, or a phenyl lower alkyl group. ] The alkylation of formula 1a can be carried out under the same reaction conditions as the acylation of formula 1b above. When converting the lower alkanoyl group of the compound of formula 1d above into a hydrogen atom, a wide range of conventional hydrolysis reaction conditions can be employed, for example, conversion is carried out in an appropriate solvent in the presence of an acid or a basic compound. Examples of the solvent include water, lower alcohols such as methanol and isopropanol, ethers such as dioxane and tetrahydrofuran, and mixed solvents thereof. Examples of acids include mineral acids such as hydrochloric acid, sulfuric acid, and hydrobromic acid, and examples of basic compounds include metal oxides such as sodium hydroxide, potassium hydroxide, and calcium hydroxide. The hydrolysis reaction normally proceeds suitably at about 0 to 150°C, and is generally completed in about 1 to 15 hours. In addition, when converting the lower alkyl group, lower alkenyl group, or phenyl lower alkyl group of the compound of formula 1e above into a hydrogen atom, for example, hydrobromic acid,
Lewis acids such as boron trichloride, boron tribromide, aluminum chloride, tin chloride, alkylthiols/
_BF3・O _{( C2H5} ) ₂ , _BCl3・S( _CH3 ) ₂ , _BBr3・S
(CH ₃ ) ₂ , LiSR ³ (R ³ is a lower alkyl group),
The compound of formula 1e may be treated in a suitable solvent in the presence of a reagent such as (CH ₃ ) ₃ SiI or (CH ₃ ) ₃ SiBr. The above reagents should be present in at least a catalytic amount in the reaction system, usually in a catalytic amount.
It is preferable that the compound be present in an amount of equimolar to 10 times the molar amount of compound 1e. As the solvent, a wide variety of solvents used in the above-mentioned hydrolysis reactions can be used. This process is usually −
It is carried out at about 78 to 50°C for about 0.5 to 24 hours. In the case of a compound in which R ⁴ represents a benzyl group among the compounds of formula 1e, a conventional debenzylation reaction such as catalytic reduction can be applied. This catalytic reduction is carried out, for example, in a solvent similar to that used in the above hydrolysis reaction, such as palladium-carbon, palladium-
The reaction is carried out in the presence of a catalytic reduction catalyst such as black or platinum black at 0°C to around room temperature for about 0.5 to 5 hours. The catalytic reduction catalyst is usually 10-50wt for the compound of formula 1e.
% is used. In these elimination reactions, selective elimination can be carried out by appropriately selecting the type of reagent used, the amount used, the reaction temperature, etc. For example, in a compound of formula 1e in which R ⁴ represents a lower alkyl group, 1 to 6 times the mole of boron trichloride is present in the reaction system, and the treatment is carried out at about -20°C to room temperature, or If 1 to 3 times the mole of boron tribromide is present and treated at about -78 to -50℃,
9-position or 2,9 of dibenzo-p-dioxin skeleton
The -30
If treated at ~0℃, the 4,7,9 of the skeleton
2, 7, 9 or 2, 4, 9 positions, and furthermore, in the presence of 3 to 6 times the mole of boron tribromide in the reaction system, the treatment is carried out at about 0°C to room temperature. If so, the 2, 4, 7, 9 positions or 2,
The 4, 9, 3', and 5' positions can be selectively de-lower alkylated. Further, compounds of formula 1b having different substituents on the dibenzo-p-dioxin skeleton can be easily produced from compounds of formula 1a by appropriately combining the above acylation, alkylation, hydrolysis, dealkylation, etc. The compound of the present invention thus obtained can be subjected to various isolation and purification operations such as solvent distillation, solvent extraction, precipitation,
It is easily isolated and purified from the reaction mixture by recrystallization, column chromatography, preparative chromatography, high performance liquid chromatography, etc. Among the compounds of general formula (1) thus obtained, compounds having an acidic group include, for example, sodium hydroxide,
It can easily form salts by reacting with strong basic compounds such as potassium hydroxide and calcium hydroxide, and the present invention also includes salts of such dibenzo-p-dioxin derivatives. When using the compound of general formula (1) and its salt as a prophylactic or therapeutic agent for thrombosis, it is usually used in the form of a general pharmaceutical preparation. The formulation is prepared using commonly used diluents or excipients such as fillers, fillers, binders, wetting agents, disintegrants, surfactants, and lubricants. Various forms of this pharmaceutical preparation can be selected depending on the therapeutic purpose, and representative examples include tablets, pills, powders, solutions, suspensions, emulsions, granules, capsules, suppositories, and injections ( liquid agents, suspension agents, etc.). When forming tablets, a wide variety of carriers conventionally known in this field can be used, such as lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid, etc. Excipients, water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, binders such as polyvinylpyrrolidone, dry starch, sodium alginate, agar powder, laminaran powder , disintegrants such as sodium bicarbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, starch, lactose, disintegration inhibitors such as sucrose, stearin, cocoa butter, hydrogenated oil, etc. absorption enhancers such as grade ammonium bases and sodium lauryl sulfate, humectants such as glycerin and starch, adsorbents such as starch, lactose, kaolin, bentonite, and colloidal silicic acid, purified talc, stearate, boric acid powder, Examples include lubricants such as polyethylene glycol. Furthermore, tablets may be coated with conventional coatings, such as sugar-coated tablets, gelatin-coated tablets,
The tablets can be enteric-coated, film-coated, double-layered, or multilayered. When forming into a pill form, a wide variety of carriers conventionally known in this field can be used, such as excipients such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, kaolin, talc, gum arabic powder, etc. , tragacanth powder, gelatin, binders such as ethanol,
Examples include disintegrants such as laminaran and agar. When forming into a suppository, a wide variety of conventionally known carriers can be used, such as polyethylene glycol, cacao butter, higher alcohols,
Examples include higher alcohol esters, gelatin, and semi-synthetic glycerides. When prepared as injectables, solutions and suspensions are preferably sterile and isotonic with blood;
In forming these solutions, emulsions and suspensions, any diluent commonly used in this field can be used, such as water,
Examples include ethyl alcohol, propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, and polyoxyethylene sorbitan fatty acid esters. In this case, a sufficient amount of salt, glucose, or glycerin may be included in the drug to prepare an isotonic solution, and usual solubilizing agents, buffers, soothing agents, etc. may be added. Good too. Furthermore, colorants, preservatives, perfumes, flavoring agents, sweeteners, and other pharmaceuticals may be included in the drug, if necessary. The amount of the compound of general formula (1) to be contained in the drug of the present invention is not particularly limited and can be selected within a wide range, but is usually 0.1 to 70% by weight of the total composition, preferably
It is 0.5-30% by weight. There are no particular restrictions on the method of administering the drug of the present invention;
Various formulation forms, patient age, gender and other conditions,
It is administered in a manner that depends on the severity of the disease. For example, tablets, pills, solutions, suspensions, emulsions, granules and capsules are administered orally. In the case of injections, they are administered intravenously alone or mixed with regular fluids such as glucose and amino acids.
Furthermore, it may be administered alone intramuscularly, intradermally, subcutaneously, or intraperitoneally as necessary. Suppositories are administered rectally. The dosage of the drug of the present invention is appropriately selected depending on the usage, patient's age, sex and other conditions, degree of disease, etc., but the amount of the compound of general formula (1), which is the active ingredient, is usually 1 kg of body weight per day. The amount is preferably about 0.05 to 100 mg per serving. The results of pharmacological tests on the compounds of the present invention are shown below. Test compound No. 1 1-(3,5-dihydroxyphenoxy)-
2,4,7,9-tetrahydroxy-dibenzo-p-dioxin 2 1-(3,5-dihydroxyphenoxy)-
2,4,7-trihydroxy-9-methoxy-
Dibenzo-p-dioxin 3 1-(3,5-dimethoxyphenoxy)-2,
4,7,9-tetrahydroxy-dibenzo-p
-Dioxine 4 1-(3,5-dihydroxyphenoxy)-
2,4,9-trihydroxy-7-methoxy-
Dibenzo-p-dioxin 5 1-(3,5-dimethoxyphenoxy)-2-
Methoxy-4,7,9-trihydroxy-dibenzo-p-dioxin 6 1-(3,5-dimethoxyphenoxy)-2,
7,9-trihydroxy-4-methoxy-dibenzo-p-dioxin [Pharmacological test 1] As a plasmin inhibitor, it was obtained from human plasma by the method of H. Rinderknecht et al. [Biochem.Med., 14 , 162 (1975)]. The prepared alpha 2 macroglobulin was used. Alpha 2 Macroglobulin 17μg 0.1M
10 of the test compound at various concentrations in 0.3 ml of 0.05 M Tris-HCl buffer (PH = 7.4) containing sodium chloride.
Mix with 0.1 ml of % methanol aqueous solution and incubate at 20°C under 37°C.
Hold for minutes. Next, 0.1 ml of 10 μg/ml bovine trypsin (manufactured by Sigma Chemical Co., Ltd., Type) was added to the above mixture, and this was kept at 37° C. for 2 minutes. 0.5 ml of the above buffer containing 2% protamine sulfate (CradeX, manufactured by Sigma Chemical Co.) was added, and the mixture was left to stand for an additional 30 minutes. 18
The reaction was stopped by adding 3 ml of % trichloroacetic acid aqueous solution, left for 1 hour, centrifuged, 50μ of supernatant was taken into a test tube, and 4 ml of 1.5N sodium hydroxide aqueous solution of 0.01% 8-hydroxyquinoline and 0.1%
1 ml of bromosuccinimide aqueous solution was added and stirred to develop a color, and the absorbance at 500 nm was measured. Plasmin inhibitor activity inhibition rate (%) was calculated using the following formula. Inhibition rate (%) = C-B/A-B x 100 A: Absorbance when alpha 2 macroglobulin and the test compound are not included B: Absorbance when the test compound is not included and alpha 2 macroglobulin is included Absorbance C: Absorbance when alpha 2 macroglobulin and test compound are included Table 1 shows the results of determining the concentration of the test compound at which the inhibition rate determined above is 50% (50% inhibition concentration). [Table] [Pharmacological test 2] Alpha-2 as a plasmin inhibitor
A plasmin inhibitor was used. Alpha-2 plasmin inhibitor was obtained from human plasma using the method of B.Wiman et al. [Eur.J.
Biochem., 78 , 19 (1977)], and human plasmin was prepared by the method of DGDoutsch et al. [Science, 170 , 1095] from human plasma.
(1970)] was prepared by the method of M. Moroi et al. [J. Biol, Chem.,
251, 5956 (1976)] with urokinase-conjugated sepharose and used for testing. Alpha 2 plasmin inhibitor 3μg
Contains 0.06M Tris containing 0.09M Sodium Chloride
Add 0.1 ml of test compounds at various concentrations dissolved in 10% methanol to 0.7 ml of hydrochloric acid buffer (PH = 7.4),
It was held at 37°C for 20 minutes. Next, 0.1 ml of a 0.5 casein unit/ml human plasmin solution dissolved in 0.1 M sodium phosphate buffer (PH = 7.4) containing 25% glycerin was added, and after heating at 37°C for 30 seconds, S at a concentration of 3 mM was added as a substrate. -2251 aqueous solution (H-D-
0.1 ml of Val-L-Leu-L-Lys-p-nitroanilide (Daiichi Kagaku) was added thereto, and the mixture was further reacted at 37°C for 3 minutes.
The reaction was stopped by adding 0.1 ml of 50% acetic acid aqueous solution. The absorbance of the reaction solution at 405 nm was measured, and the inhibition rate (%) of plasmin inhibitor activity was calculated using the following formula. Inhibition rate (%) = C-B/A-B x 100 A: Absorbance when alpha 2/plasmin inhibitor and test compound are not included B: When alpha 2/plasmin inhibitor alone is included without the test compound Absorbance C: Absorbance when alpha-2/plasmin inhibitor and test compound are included The inhibition rate of the test compound against alpha-2/plasmin inhibitor determined by the above method is 50
% concentration (50% inhibition concentration) is shown in Table 2 below. [Table] Examples and formulation examples are listed below. Example 1 (1) Fresh Kurome (collected in Irino, Kochi Prefecture) 600Kg
was extracted with methanol at room temperature. The extract was concentrated under reduced pressure to obtain a gummy primary extract.
This was extracted repeatedly with ethyl acetate-water (1:1/) until the upper layer became colorless, and the obtained upper layer was concentrated under reduced pressure to obtain 5.7 kg of a second extract. (2) 1.7 kg of the secondary extract obtained above was mixed with 3.4 kg of Celite (manufactured by Johns Manvills) and dried under reduced pressure. The obtained solid was finely ground and packed into a glass column, and benzene (18), methylene chloride (36), and ethyl ether (54)
After sequential elution with methanol. Sephadex 552g eluted with ethyl ether
It was subjected to LH-20 (3.5Kg) column chromatography and eluted with acetone (15) to obtain fractions of 2000ml each. The crude crystals obtained by concentrating fraction No. 4 under reduced pressure are recrystallized from hot water to obtain 1-
(3,5-dihydroxyphenoxy)-2,4,
40 g of 7,9-tetrahydroxy-dibenzo-p-dioxin [compound of formula 1a] was obtained. The physical properties of the obtained compound are shown below. Melting point 243-244℃ Colorless plate crystal λ ^MeOH _nax : 230 (ε=30000) 290 (ε=3160) nm IR (ν ^KBr _nax ): 3250, 1605, 1475, 1370, 1260,
1200, 1160, 1140, 1110, 1083, 1040,
1010, 805 cm ^-1 high-resolution mass spectrum Actual value 372.0460 Theoretical value 372.0481 (as C ₁₈ H ₁₂ O ₉ ) PMR (200 MHz, DMSO−d ₆ , ppm): 5.73
(2H, d, J=2.1), 5.81 (1H, d, J=
2.6), 5.82 (1H, t, J=2.1), 5.97 (1H,
d, J=2.6), 6.16 (1H, s), 9.15 (2H,
s), 9.18 (1H, s), 9.19 (1H, s), 9.46
(1H, s), 9.51 (1H, s) CMR (100MHz, DMSO-d ₆ , ppm): 94.0,
96.3, 98.4, 98.7, 122.6, 122.9, 123.5,
137.3, 141.9, 142.7, 145.9, 146.1,
153.0, 158.7, 160.4 Example 2 200 mg of 1-(3,5-dihydroxyphenoxy)-2,4,7,9-tetrahydroxy-dibenzo-p-dioxin obtained in Example 1 was dissolved in pyridine 2.
ml and 1 ml of acetic anhydride and reacted at room temperature for 20 hours. The reaction mixture was added to ice water, and the resulting precipitate was separated and recrystallized from benzene-methanol to give 1-(3,5-diacetyloxyphenoxy).
280 mg of -2,4,7,9-tetraacetyloxy-dibenzo-p-dioxin was obtained. Melting point 211-212.5℃ Colorless prismatic crystals EIMS (m/z): 624 (M ⁺ ), 582, 540, 498,
456, 414, 372, 69, 43 IR (ν ^KBr _nax ): 2900, 2850, 1750, 1595, 1490,
1435, 1360, 1175, 1120, 1100, 1010,
875cm ^-1 PMR (400MNz, CDCl ₃ , ppm): 1.96 (3H,
s), 2.13 (3H, s), 2.24 (3H, s), 2.26
(6H, s), 2.34 (3H, s), 6.50 (1H, d,
J=2.6), 6.58 (2H, d, J=2.0), 6.63
(1H, d, J=2.6), 6.66 (1H, s), 6.73
(1H, t, J=2.0) Example 3 1-(3,5-dihydroxyphenoxy)-2,
4,7,9-tetrahydroxy-dibenzo-p-
A mixture of 500 mg of dioxin, 1.3 g of anhydrous potassium carbonate, 3 ml of methyl iodide and 10 ml of anhydrous DMF was stirred at room temperature for one day. The reaction solution was acidified by adding a 2N aqueous hydrochloric acid solution, and then extracted with ethyl acetate. The extracted layer was thoroughly washed with water and saturated brine, dried (magnesium sulfate), and the solvent was distilled off under reduced pressure to obtain a red oil. This oil was purified by silica gel (10 g: Wako Gel C-200) column chromatography, and the resulting crystals were recrystallized from ether-methylene chloride to produce 1-(3,5-dimethoxy)-2,4,7,
9-Tetramethoxy-dibenzo-p-dioxin
Obtained 510 mg. Melting point 171-172℃ Colorless prismatic crystals IR (ν ^KBr _nax ): 2925, 2830, 1590, 1500, 1450,
1370, 1200, 1110, 1050, 970, 910, 810,
785cm ^-1 EIMS (m/z): 456 (M ⁺ ) PMR (400MHz, CDCl ₃ , ppm): 3.66 (3H,
s), 3.71 (9H, s), 3.77 (3H, s), 3.92
(3H, s), 6.09 (1H, d, J=2.9), 6.13
(1H, t, J=2.1), 6.17 (2H, d, J=
2.1), 6.26 (1H, s) Example 4 1-(3,5-dihydroxyphenoxy)-2,
4,7,9-tetrahydroxy-dibenzo-p-
To a solution of 200 mg of dioxin in 5 ml of DMF was added 216 mg of NaH (50% oil) at 0°C under a nitrogen stream, and the reaction mixture was stirred at room temperature for 20 minutes. After dropping 0.5 ml of benzyl bromide and further stirring at room temperature for 5 hours,
Ice water was added, acidified with 2N aqueous hydrochloric acid, and extracted with ether. The ether layer was washed with water and saturated brine, dried (magnesium sulfate), and then evaporated under reduced pressure to obtain an oil. This oil was subjected to silica gel (5 g) column chromatography, and the crystals obtained by elution with benzene were recrystallized from ether.
(3,5-dibenzyloxyphenoxy)-2,
4,7,9-tetrabenzyloxy-dibenzo-
400 mg of p-dioxin was obtained. Melting point 141-142°C Colorless prismatic crystals EIMS (m/z): 912 (M ⁺ ) Example 5 1-(3,5-dihydroxyphenoxy)-2,
4,7,9-tetrahydroxy-dibenzo-p-
216 mg of NaH (50% oil) was added to a solution of 200 mg of dioxin in 5 ml of DMF under a nitrogen stream at 0°C, and the mixture was heated at room temperature for 20
Stir for a minute. Administer 484 mg of allyl bromide,
After further stirring at room temperature for 5 hours, ice water was added and 2N-
The reaction solution was acidified with aqueous hydrochloric acid and extracted with ether. After washing the ether layer with water and saturated brine and drying (magnesium sulfate), the ether was removed under reduced pressure to obtain amorphous 1-(3,5-diallyloxyphenoxy)-2,4,7,9. 300 mg of -tetraallyloxy-dibenzo-p-dioxin was obtained. IR (ν ^film _nax ): 3100, 3025, 2950, 2900, 1605,
1510, 1460, 1425, 1360, 1245, 1210,
1130, 1050, 990, 930, 820, 730 cm ^-1 EIMS (m/z): 612 (M ⁺ ) Example 6 1-(3,5-dihydroxyphenoxy)-2,
4,7,9-tetrahydroxy-dibenzo-p-
To a solution of 100 mg of dioxine in 5 ml of methanol was added dropwise 15 ml of a diazomethane-ether solution prepared from 200 mg of nitrosomethylurea at 0°C, and the mixture was left at room temperature overnight. The solvent was distilled off under reduced pressure to obtain a viscous liquid. This was subjected to silica gel (10 g) column chromatography and extracted with a 10% methanol chloroform solution to successively isolate the following compounds. (a) 1-(3,5-dihydroxyphenoxy)-
2,4,9-trimethoxy-7-hydroxy-
Dibenzo-p-dioxin Colorless needle crystals (chloroform-methanol) Yield 30mg Melting point 267-268℃ EIMS (m/z): 414 (M ⁺ ), 382, 315, 260,
217, 123, 69, 53 PMR (400MHz, _CDCl3 /DMSO-d ₆ ppm):
3.65 (3H, s), 3.75 (3H, s), 3.89 (3H,
s), 6.00 (2H, dJ=2.0), 6.05 (1H, t,
J=2.0), 6.07 (1H, d, J=2.6), 6.15
(1H, d, J=2.6), 6.24 (1H, s), 8.25
(2H, s), 8.60 (1H, s) (b) 1-(3,5-dihydroxyphenoxy)-
2,7-dihydroxy-4,9-dimethoxy-
Dibenzo-p-dioxin Colorless prismatic crystals (chloroform-methanol) Yield 43mg Melting point 287-288℃ EIMS (m/z): 400 (M ⁺ ), 367, 276, 246,
203, 111, 69 PMR (400MHz, DMSO-d ₆ ppm): 3.62 (3H,
s), 3.76 (3H, s), 5.74 (2H, d, J=
2.2), 5.83 (1H, t, J=2.2), 5.96 (1H,
d, J=2.4), 6.29 (1H, s), 6.08 (1H,
d, J=2.4), 9.18 (2H, s), 9.45 (2H,
s) (c) 1-(3,5-dihydroxyphenoxy)-
2,4,7-trihydroxy-9-methoxy-
Dibenzo-p-dioxin Colorless prismatic crystals (methanol-water) Yield 30mg Melting point 196-197℃ EIMS (m/z): 386 (M ⁺ ), 353, 232, 193,
69 PMR (400MHz, DMSO-d ₆ ppm): 3.62 (3H,
s), 5.73 (2H, d, J = 2.0), 5.81 (1H,
t, J = 2.0), 5.96 (1H, d, J = 2.7),
6.07 (1H, d, J = 2.7), 6.16 (1H, s),
9.15 (2H, s), 9.24 (1H, s), 9.43 (1H,
s), 9.50 (1H, s) Example 7 1-(3,5-dihydroxyphenoxy)-2,
4,7,9-tetrahydroxy-dibenzo-p-
Dioxine 300mg anhydrous THF 10ml and anhydrous
To 3 ml of HMPA solution, 0.7 ml of a 1,39M n-butyllithium solution in n-hexane was added dropwise at -78°C under a nitrogen stream. After dropping, stir at -78℃ for 10 minutes,
A solution of 164 mg of benzyl bromide in 1 ml of THF was added dropwise, and the mixture was further stirred at -78°C for 30 minutes and then at 0°C for 1 hour. Take a part of this reaction solution, 1-(3,
5-dihydroxyphenoxy)-2,4-dibenzyloxy-7,9-dihydroxy-dibenzo-
Production of p-dioxin was confirmed. [EIMS(m/
z): 552 (M ⁺ ), 461, 370, 444, 262, 91] NaH (50% oil) was added to the reaction solution obtained above.
200 mg was added at 0°C, and after stirring for 10 minutes, 0.8 ml of methyl iodide was added dropwise. After the completion of the dropwise addition, continue at room temperature for 5 minutes.
The mixture was stirred for an hour, added with ice water, and extracted with ether.
The ether layer was washed with water and saturated brine, dried (magnesium sulfate), and then distilled off under reduced pressure to form an oil.
Obtained 577mg. Take some of this oil and 1-
(3,5-dimethoxyphenoxy)-2,4-dibenzyloxy-7,9-dimethoxy-dibenzo-
Production of p-dioxin was confirmed. [EIMS(m/
z): 608 (M ⁺ ), 594, 580, 566, 517, 426.91〕 Add this oil to 10 ml of ethanol and 5 ml of ethyl acetate.
and 3 drops of acetic acid, and hydrogenated under 10% Pd-C (20 mg) catalyst. After separating the catalyst, the oil obtained by distilling off the solvent was subjected to Bio-Bead SX-12 column chromatography and eluted with benzene to obtain 1-(3,5-
60 mg of dimethoxyphenoxy-2,4-dihydroxy-7,9-dimethoxy-dibenzo-p-dioxin was obtained. Colorless amorphous EIMS (m/z): 428 (M ⁺ ), 397, 291, 260,
214, 170, 125, 95, 53 PMR (400MHz, _CDCl3 , ppm): 3.65 (3H,
s), 3.73 (3H, s), 3.75 (6H, s), 6.10
(1H, d, J=2.7), 6.11 (1H, d, J=
2.7), 6.17 (1H, t, J = 2.1), 6.21 (2H,
d, J=2.1), 6.32 (1H, s), 5.15 (1H,
s), 5.21 (1H, s) Example 8 1-(3,5-dimethoxyphenoxy)-2,
4,7,9-tetramethoxy-dibenzo-p-dioxin 60mg in dichloromethane 2ml solution at 1.55M
After dropping 0.26 ml of a dichloromethane solution of -BC ₃ at -78°C, the mixture was stirred at the same temperature for 3 hours and then at room temperature for 3 hours. After adding water and acidifying with 2N hydrochloric acid solution,
Extraction was performed with dichloromethane, and the solvent was distilled off under reduced pressure to obtain 50 mg of oil. This oil was subjected to medium pressure column chromatography (Robar Lichroprep Si-60, TypA) and chloroform-ethyl acetate (9:1V/V)
The following compounds were successively isolated by elution. (a) 1-(3,5-dimethoxyphenoxy)-2,
4,7-Trimethoxy-9-hydroxy-dibenzo-p-dioxin Colorless prismatic crystals (ether-dichloromethane) Yield 30 mg Melting point 196-197℃ EIMS (m/z): 442 (M ⁺ ), 380, 353, 288,
207, 151, 69 PMR (400MHz, _CDCl3 , ppm): 3.69 (3H,
s), 3.75 (6H, s), 3.80 (3H, s) 3.93
(3H, s), 4.93 (1H, s), 6.12 (1H, d,
J=2.8), 6.3 (2H, d, J=2.2), 6.16
(1H, t, J=2.2) 6.17 (1H, d, J=
2.8) (b) 1-(3,5-dimethoxyphenoxy)-
2,9-dihydroxy-4,7-dimethoxy-
Dibenzo-p-dioxy Amorphous Yield 15 mg EIMS (m/z): 428 (M ⁺ ), 385, 274, 214,
151, 69 PMR (400MHz, CDCl ₃ ppm): 3.69 (3H, s),
3.75 (6H, s), 3.89 (3H, s), 6.16 (1H,
d, J=2.8), 6.11 (1H, d, J=2.8),
6.18 (2H, d, J = 2.2), 6.20 (1H, t,
j=2.2), 6.35 (1H, s), 4.78 (1H, s),
5.22 (1H, s) Example 9 1-(3,5-dimethoxyphenoxy)-2,
A solution of 100 mg of 4,7,9-tetramethoxy-dibenzo-p-dioxin in 4 ml of dichloromethane
0.838M-BBr ₃ dichloromethane solution 1.57ml
The mixture was added dropwise at °C, and stirred at the same temperature for 1 hour and then at room temperature for 20 hours. After adding water and acidifying with 2N hydrochloric acid,
Extraction was performed with ethyl acetate, and the solvent was distilled off under reduced pressure to obtain 70 mg of oil. This oil was subjected to silica gel 5g column chromatography and eluted with methanol-chloroform (1:9V:V) to successively isolate the following compounds. (a) 1-(3,5-dimethoxyphenoxy)-2,
4,7,9-tetrahydroxy-dibenzo-p
-Dioxine Amorphous Yield 27mg EIMS (m/z): 400 (M ⁺ ), 352, 325, 287,
260, 245, 193, 154, 122, 69 PMR (400NHz, DMSO−d ₆ , ppm): 3.68
(6H, s), 5.80 (1H, d, J=2.7), 5.96
(1H, d, J=2.7), 6.00 (2H, d, J=
2.2), 6.14 (1H, t, J=2.2), 6.16 (1H,
s), 9.19 (1H, s), 9.27 (1H, s), 9.50
(1H, s), 9.51 (1H, s) (b) 1-(3,5-dihydroxyphenoxy)-
2,4,9-trihydroxy-7-methoxy-
Dibenzo-p-dioxin Amorphous Yield 25 mg EIMS (m/z): 386 (M ⁺ ), 325, 287, 246,
123, 95, 69 PMR (400MHz, CDCl ₃ +10% DMSO−
d ₆ ppm): 3.71 (3H, s), 5.99 (1H, d,
J=2.7), 6.03 (2H, d, J=2.2), 6.03
(1H, d, J=2.7), 6.10 (1H, t, J=
2.2) Example 10 1-(3,5-dihydroxyphenoxy)-2,
10 mg of 4,7-trihydroxy-9-methoxy-dibenzo-p-dioxin was mixed with 0.5 ml of pyridine and 0.1 ml of acetic anhydride, and reacted for 5 hours. The reaction solution was added to ice water, and the resulting precipitate was separated and recrystallized from methanol to give 1-(3,5-diacetyloxyphenoxy)-2,4,7-triacetyloxy-9-methoxy- 11 mg of dibenzo-p-dioxin was obtained. Colorless prismatic crystals Melting point 205-207℃ EIMS (m/z): 596 (M ⁺ ), 554, 512, 470,
428, 409, 386, 357, 315, 278, 232,
191, 137, 109, 69, 43 PMR (400MHz, _CDCl3 , ppm): 2.09 (3H,
s), 2.25 (9H, s), 2.32 (3H, s), 3.66
(3H, s), 6.28 (1H, d, J=2.5), 6.32
(1H, d, J=2.5), 6.62 (1H, s), 6.64
(2H, d, J=2.0), 6.67 (1H, t, J=
2.0) Example 11 1-(3,5-dihydroxyphenoxy)-2,
15 mg of 7-dihydroxy-4,7-dimethoxy-dibenzo-p-dioxin, 0.8 ml of pyridine and 0.2 ml of acetic anhydride were reacted for 5 hours. The reaction solution was added to ice water, and the resulting precipitate was separated and recrystallized from methanol to give 1-(3,5-diacetyloxyphenoxy)-2,7-diacetyloxy-4,7.
15 mg of -dimethoxy-dibenzo-p-dioxin was obtained. Colorless prismatic crystals Melting point 201-202℃ EIMS (m/z): 568 (M ⁺ ), 526, 484, 442,
400, 371, 315, 292, 246, 203, 177,
123, 69, 43 PMR (400NHz, _CDCl3 , ppm): 2.12 (3H,
s), 2.25 (6H, s), 2.29 (3H, s), 3.66
(3H, s), 3.86 (3H, s), 6.28 (1H, d,
J=2.5), 6.40 (1H, d, J=2.5), 6.42
(1H, s), 6.61 (2H, s), 6.66 (1H, t,
J=2.0) Example 12 1-(3,5-dimethoxyphenoxy)-2,
0.838 M in a solution of 100 mg of 4,7,9-tetramethoxy-dibenzo-p-dioxin in 4 ml of methylene chloride.
A solution of 0.79 ml of -BBr ₃ in dichloromethane was added dropwise at -25°C. After stirring at -25°C for 2 hours, water was added and the mixture was extracted with ethyl acetate. The organic layer was dried over magnesium sulfate and then distilled off under reduced pressure to obtain 60 mg of oil. Dissolve this in 0.1 ml of methanol and perform high performance liquid chromatography (Hypersil-ODS, 10 x 300 mm, solvent 50 V/V% water-methanol, flow rate 3 ml/min).
The following compound was obtained. (a) 1-(3,5-dimethoxyphenoxy)-2,
7,9-trihydroxy-4-methoxy-dibenzo-p-dioxin Colorless powder The above HPLC retention time 23 minutes Yield 30 mg EIMS (m/z): 414 (M ⁺ ), 366, 339, 301,
274, 207, 165, 122, 69, 53, 28 PMR (400MHz, _CDCl3 , ppm): 3.75 (6H,
s), 3.88 (3H, s), 6.04 (1H, d, J=
2.7), 6.08 (1H, d, J = 2.7), 6.17 (2H,
d, J = 2, 1), 6.20 (1H, t, J =
2.1), 6.34 (1H, s) (b) 1-(3,5-dimethoxyphenoxy)-2,
4,9-trihydroxy-7-methoxy-dibenzo-p-dioxin Colorless powder HPLC retention time 25 minutes Yield 5 mg EIMS (m/z): 414 (M ⁺ ), 371, 316, 260,
207, 177, 123, 95, 69, 53, 28 (c) 1-(3,5-dimethoxyphenoxy)-2-
Methoxy-4,7,9-trihydroxy-dibenzo-p-dioxin Colorless powder HPLC retention time 28 minutes Yield 45 mg EIMS (m/z): 414 (M ⁺ ), 383, 339, 274,
193, 151, 122, 69, 39 PMR (400MHz, _CDCl3 , ppm): 3.75 (9H,
s), 6.02 (1H, d, J=2.7), 6.08 (1H,
d, J = 2.7), 6.15 (1H, t, J = 2.1),
6.13 (1H, d, J = 2.1), 6.32 (1H, s) Reference example 1 Compound of the present invention (compound of Example 1) 500 mg Glucose 250 mg Distilled water for injection Appropriate amount Total amount 5 ml Add the compound of the present invention to distilled water for injection After dissolving the sodium salt and glucose, pour into a 5 ml ampoule. After purging with nitrogen, autoclave sterilization at 121°C for 15 minutes to obtain an injection. Reference Example 2 Compound of the present invention (compound of Example 2) 150.0g Citric acid 1.0g Lactose 33.5g Calcium phosphate 70.0g Pluronic F-68 30.0g Sodium lauryl sulfate 15.0g Polyvinylpyrrolidone 15.0g Polyethylene glycol (Carbovacs) 1500) 4.5g Polyethylene glycol (Carbowax 6000) 45.0g Cornstarch 30.0g Dry sodium lauryl sulfate 3.0g Dry magnesium stearate 3.0g Ethanol Appropriate amount Compound of the present invention, citric acid, lactose, dicalcium phosphate, Prone F-68 and sodium Mix lauryl sulfate. The above mixture is sieved through a No. 60 screen and wet granulated with an alcoholic solution consisting of polyvinylpyrrolidone, Carbowax 1500 and 6000. Add alcohol if necessary to make the powder into a pasty mass. Add cornstarch and continue mixing until uniform particles are formed. Pass it through a No. 10 screen, put it in a tray and put it in an oven at 100℃ for 12~
Dry for 14 hours. The dry particles are sieved through a No. 16 screen, mixed with dry sodium uralyl sulfate and dry magnesium stearate, and compressed into the desired shape using a tablet machine. The core is treated with varnish and sprinkled with talc to prevent moisture absorption. A subbing layer is applied around the core. Apply varnish enough times for internal use. Further subbing layers and smooth coatings are applied to make the tablet perfectly round and smooth. Pigmented coatings are applied until the desired shade is obtained. After drying, the coated tablets are polished to a uniform gloss.

[Brief explanation of drawings]

Figure 1 shows the PMR of the compound obtained in Example 1.
It is a spectrum diagram. FIG. 2 is a CMR spectrum diagram of the compound obtained in Example 1. FIG. 3 is a PMR spectrum diagram of the compound obtained in Example 3. FIG. 4 is a PMR spectrum diagram of the compound obtained in Example 6a. FIG. 5 is a PMR spectrum diagram of the compound obtained in Example 6b. Figure 6 shows the compound obtained in Example 6c.
It is a PMR spectrum diagram. Figure 7 shows Example 7.
FIG. 2 is a PMR spectrum diagram of a compound obtained in FIG.
FIG. 8 is a PMR spectrum diagram of the compound obtained in Example 8a. FIG. 9 is a PMR spectrum diagram of the compound obtained in Example 8b. 10th
The figure is a PMR spectrum diagram of the compound obtained in Example 9a. FIG. 11 is a PMR spectrum diagram of the compound obtained in Example 9b. FIG. 12 is a PMR spectrum diagram of the compound obtained in Example 12a. FIG. 13 is a PMR spectrum diagram of the compound obtained in Example 12c.

Claims (1)

[Claims] 1. General formula [In the formula, R is the same or different and represents a hydrogen atom, a lower alkyl group, a lower alkenyl group, a phenyl lower alkyl group, or a lower alkanoyl group. ] A dibenzo-p-dioxin derivative represented by these and its salt.